荻草谷网蚜体内杀虫剂抗性相关基因的分析及其功能验证
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Citation：张孟元,胡贵磊,蒋宇泰,胡帅飞,艾春旺,赵卫忠,张百重.荻草谷网蚜体内杀虫剂抗性相关基因的分析及其功能验证.Journal of Plant Protection,2023,50(2):446-458
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Author NameAffiliationE-mail
Zhang Mengyuan College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Hu Guilei College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Jiang Yutai College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Hu Shuaifei College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Ai Chunwang College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Zhao Weizhong College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Zhang Baizhong College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China baizhongzhang@hist.edu.cn 
中文摘要:为了解荻草谷网蚜Sitobion miscanthi体内与杀虫剂靶标和代谢抗性相关的基因信息及其在不同发育阶段的表达情况,通过转录组测序选出不同发育阶段差异性表达的抗药性相关基因,并采用荧光定量PCR和RNA干扰技术对选取的5个抗药性相关基因进行功能验证。结果显示,共获得与杀虫剂靶标和代谢相关基因374个,其中在3龄若蚜体内上调表达基因有129个。CAT-3、CYP6A13-2、CYP4g15-2和nAChR α2-3基因在3龄若蚜、无翅成蚜和有翅成蚜体内的表达量比在1龄若蚜体内的表达量均显著上调。GST1-1基因在3龄若蚜和有翅成蚜体内的表达量比在1龄若蚜体内的表达量显著上调。荻草谷网蚜3龄若虫经吡虫啉处理后,CAT-3、CYP6A13-2、CYP4g15-2和GST1-1的表达量均显著上调,而nAChR α2-3的表达量显著下调。沉默CAT-3、CYP6A13-2、CYP4g15-2及GST1-1后蚜虫对吡虫啉的敏感度增加,而沉默nAChRα2-3后蚜虫对吡虫啉的敏感度降低,表明CAT-3、CYP6A13-2、CYP4g15-2、nAChR α2-3和GST1-1这5个基因均可能与荻草谷网蚜对吡虫啉的抗性相关。
中文关键词:荻草谷网蚜  转录组  杀虫剂靶标相关基因  解毒酶相关基因  荧光定量PCR  RNA干扰
 
Analysis and functional verification of the genes related to insecticide resistance in Indian grain aphid Sitobion miscanthi
Author NameAffiliationE-mail
Zhang Mengyuan College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Hu Guilei College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Jiang Yutai College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Hu Shuaifei College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Ai Chunwang College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Zhao Weizhong College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China  
Zhang Baizhong College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China baizhongzhang@hist.edu.cn 
Abstract:In order to understand the genes related to insecticide targets and metabolic resistance in Indian grain aphid Sitobion miscanthi and their expression changes at different developmental stages, the genes related to insecticide resistance and differentially expressed among different developmental stages based on transcriptome sequencing were selected, and the functions of five selected genes were verified with fluorescence quantitative real-time PCR and RNA interference techniques. The results showed that 374 genes related to insecticide targets and metabolism were obtained, and 129 genes were up-regulated in the 3rd-instar nymph. The expressions of CAT-3, CYP6A13-2, CYP4g15-2 and nAChRα2-3 were significantly up-regulated in the 3rd-instar nymph, apterous adult and winged adult compared with in the 1st-instar nymph. The expression of GST1-1 was significantly up-regulated in the 3rd-instar nymph and winged adult compared with in the 1st-instar nymph. In addition, the expression levels of CAT-3, CYP6A13-2, CYP4g15-2, and GST1-1 were significantly up-regulated, while nAChRα2-3 was significantly down-regulated in the 3rd-instar nymph under imidacloprid treatment. Silencing CAT-3, CYP6A13-2, CYP4g15-2 and GST1-1 resulted in increased sensitivity of aphids to imidacloprid, while silencing nAChRα2-3 resulted in decreased sensitivity of aphids to imidacloprid. These results indicated that the five selected genes are most likely related to imidacloprid resistance.
keywords:Sitobion miscanthi  transcriptome  insecticide target related genes  detoxification enzyme related genes  fluorescence quantitative real-time PCR  RNA interference
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